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Enhanced Bioactivity of the Anti-LOX-1 scFv Engineered by Multimerization Strategy

Author:
Hu, Wei, Xie, Qiuhong, Liu, Ling, Xiang, Hongyu
Source:
Applied biochemistry and biotechnology 2018 v.185 no.1 pp. 233-247
ISSN:
0273-2289
Subject:
Brevibacillus choshinensis, antigens, atherosclerosis, bioactive properties, blood serum, half life, hosts, immune response, low density lipoprotein, monoclonal antibodies, neutralization, oxidation, secretion, therapeutics
Abstract:
Single-chain variable fragment (scFv) antibodies as therapeutic agents have the potential to reduce the production cost and immunogenicity relative to monoclonal antibodies, but their monovalency and lack of a fragment crystallizable region can lead to reduced function. Multimerization is one strategy for recovering the function; however, their application is limited by the production of multimeric proteins. In our previous study, an anti-lectin-like oxidized low-density lipoprotein receptor-1 (LOX-1) scFv showed potential use in diagnosis and therapy of atherosclerotic diseases, but is limited by its inherent low antigen-binding activity. In this study, to improve the efficacy of the anti-LOX-1 scFv, we constructed the anti-LOX-1 scFv multimers by modifying the linker length between the variable domains of the scFv or by fusing the scFv with self-merization domains and expressed these scFv multimers in Brevibacillus choshinensis hosts. After optimization, all of the scFv multimers obtained efficient secretion expression. Compared with the scFv monomer, the multimers that are successfully fractionated displayed increased neutralization activity and showed elevated antigen-binding avidity, especially the tetramer, which improved the antigen avidity by two orders of magnitude. Moreover, the scFv dimer and the tetramer both displayed better stability and longer half-life in serum, which can be attractive candidates for the next-generation anti-LOX-1 therapeutic antibody.
Agid:
5936225